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文献和实验Long PCR Reagents and Guidelines
. Generic Long PCR Program for the Perkin Elmer 9600 Initial melting 94 degrees C 10-15 sec Cycles 1-15 94 degrees C 10 sec, 68 degrees C for n min (15 times) Cycles 16-30 94 degrees C 10 sec, 68 degrees C for n min +15 sec/cycle (15 times)
High Efficiency Yeast Transformation (Gietz Long Protocol)
and remove the LiAc with a micropipette. 11. The basic "transformation mix" consists of: 240 µl PEG (50% w/v) 36 µl 1.0 M. LiAc 50 µl SS-DNA (2.0 mg/ml) X µl Plasmid DNA (0.1 - 10 µg) 34-X µl Sterile ddH2O 360 µl TOTAL Carefully add these ingredients
Long-PCR Reagents and Guidelines
Tricine pH 8.7 (adjust pH of stock solution with KOH) 8% glycerol 1% DMSO (1 to 4% works) A 5X buffer stock containing 5% DMSO (1% final conc.) can easily be made using 1 M Tricine and 1 M KOAc stock solutions as follows: 10 ml 5X buffer 4.25 ml
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